Screening Apomixis Related Gene Of Boehmeria Tricuspis Diploid With MRNA Differential Display

Apomixis has potential applications for crop breeding. The research of apomixis genes was good to reveal its genetic mechanism and applied in practice. Megaspore mother cells period was the branch point of sexual and apomixis reproduction, and it was also the most active period of apomixis gene expression. This materials in this study were Boehmeria tricuspis diploid, measure the inflorescence length, flower count, rachis diameter, flowers diameter and others, and whole clearing techniques was used for selecting ovule at megaspore mother cells period, meanwhile, which was used for the analysis of external morphology characteristics and establishing corresponding system of external morphology index and the development of internal ovule. Through the method of mRNA differential display, the ovule gene expression differences of apomixis type and sexual type of Boehmeria tricuspis diploid at the megaspore mother cell period were analyzed. Total RNA of two reproductive type of Boehmeria tricuspis diploid were respectively extracted during the megaspore mother cell period, which were used as initial template for reverse transcription - polymerase chain reaction (RT-PCR), and then difference segments were recycled and analyzed. The differences bands were sequenced and compared by BLASTN on NCBI, and the result showed that 4 in the 14 separated differences bands were related to reproductive regulation, and these related genes segments were respectively fis gene , F - box gene , PIN9 protein gene and Ricinus hypothetical protein gene.